Abstract: Abstract Objective: To construct the luciferase reporter gene vector of cell division cycle 2 gene promoter and determine its transcriptional activity. Methods: Primers were designed based on human Cdc2 promoter sequence from UCSC software. Then Cdc2 promoter from human genome DNA was replicated. After pGL3-Basic Vector and Cdc2 promoter were digested with restriction enzymes ScaⅠand XhoⅠseparately, Cdc2 promoter was inserted into pGL3-Basic vector. The recombinant plasmid named pGL3-Cdc2-promoter was transiently co-transfected into U2OS cells with control vector pRL-SV40, then the activity of dual luciferase was detected. Results: pGL3-Cdc2- promoter was constructed successfully. Restriction analysis and sequencing proved the same as the design . Promoter activity of pGL3-Cdc2-promoter in U2OS was 32 times than that in negative control group. Conclusion: pGL3-Cdc2-promoter can be transcribed and activated in U2OS cells. This research provided an important basis for antitumor drug screening and evaluation.

Key words: cell division cycle 2, Cdc2, cyclin-dependent kinase 1, CDK1, cell cycle, promoter, dual-Luciferase